After 50-fold overdigestion with Bpu1102I, more than 80% of the DNA fragments can be ligated in a reaction mixture containing 20-40 u of T4 DNA Ligase/1μg of fragments and 10% PEG. More than 95% of these can be recut

Methylation Effects:

Dam: never overlaps — no effect

Dcm: never overlaps — no effect

CpG: may overlap — no effect.

EcoKI: never overlaps — no effect.

EcoBI: may overlap — effect not determined

Digestion of Agarose-embedded DNA:

Minimum 5units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours